TNF-α基因转染增强树突细胞迁移及CCR7受体表达

Enhancement of the Expression of Chemokine Receptor CCR7 in Dendritic Cells and Their Migration by TNF-α Gene Transfer

为探讨腺病毒介导肿瘤坏死因子 - α(TNF- α)基因转染诱导树突细胞 (DCs)趋化因子受体差异性表达 ,分别用 10 0 MOI的 Ad V- TNF- α和 Ad V- p L p A (无外源基因插入 )感染小鼠树突细胞 (m DCs) ,经流式细胞仪检测细胞表型变化 ,RNA酶保护实验检测其趋化因子受体表达 ,体内、体外细胞趋化试验分析比较其迁移差异。发现 Ad V- TNF-α感染 m DCs后 CD11b、CD40、CD86 ,ICAM- 1以及 CCR7受体表达较对照组明显增高 ,CCR2受体表达量却下调 ;体外细胞趋化试验表明 DCTNF -α对 MIP- 3β(CCR7配体 )迁移应答增强 ,体内细胞趋化试验显示 DCTNF-α淋巴结趋化迁移效率较空白对照组和 Ad V- p L p A感染组分别增高 7倍和 3倍。提示 :Ad V- TNF- α感染促使 m DCs成熟 。

To investigate the differential expression of the chemokine receptors in mouse dendritic cells (mCDs) by adenovirus-mediated TNF-α gene transfer, mDCs infected with AdV-TNF-α and AdV-pLpA (with no gene insert) at 100 MOI respectively were analyzed by flow cytometry for their phenotype change, and their chemokine receptor expression was detected by RNase protection assay; Chemotaxis assay in vitro and the migration assay in vivo for DCs were also performed to determine their chemokine receptor function. Compared with AdV-pLpA and mock-infected DCs, AdV-TNF-α-infected DCs displayed up-regulated expression of CD11b, CD40, CD86 and ICAM-1 molecules, and CCR7 receptor, down-regulated the expression of CCR2 as well. Furthermore, DC TNF-α showed enhanced migratory responses for the CCR7 ligand MIP-3β in chemotaxis assays, and with 7 and 3 folds more efficient in migration to lymph nodes than untransfected DCs and DC pLpA, respectively. It was suggested that AdV-TNF-α infection might promote the maturation of mDCs and enhance the efficiency in migration to lymph nodes.